Zhang Lei, Wen Hong-tao, Zhang Lan, Chen Kui-sheng, Zhang Yun-han
Department of Pathology, First Affiliated Hospital, Zhengzhou University, Henan Key Laboratory of Tumor Pathology, Zhengzhou 450052, China.
Zhonghua Bing Li Xue Za Zhi. 2005 Jul;34(7):402-6.
To investigate the biologic effects of Bcl-XL antisense oligodeoxynucleotide (ASODN) transfected into cultured esophageal carcinoma cells and human esophageal carcinoma xenograft in nude mice.
Cationic liposome-mediated ASODN was used to transfect esophageal carcinoma cells. RT-PCR, Western blot, MTT assay, flow cytometry and in-situ apoptosis cells detection (TUNEL detection) were used to systematically study the biological effects of the transfected cells in vitro and in vivo.
MTT assay showed that the proliferation of esophageal carcinoma cells in the ASODN group decreased significantly as compared with control (P < 0.05), along with a 57.3% inhibitory rate of Bcl-XL mRNA, a significant decrease of Bcl-XL protein and the apoptosis rates of (31.1 +/- 5.8)% and 35.0% by flow cytometry and TUNEL assay, respectively (P < 0.01, as compared with controls). The growth of human esophageal carcinoma in nude mice was also significantly inhibited in the ASODN group (P < 0.05), along with a significant decrease of Bcl-XL mRNA and protein expression, and also an enhanced apoptosis of the tumor cells in nude mice.
Bcl-XL ASODN can effectively inhibit the proliferation of esophageal carcinoma cells in vitro and the growth of the tumor in vivo. The suppression of Bcl-XL expression by ASODN may offer both a therapeutic approach and an important theoretic foundation for gene therapy against esophageal carcinoma.
研究Bcl-XL反义寡脱氧核苷酸(ASODN)转染培养的食管癌细胞及人食管癌裸鼠移植瘤后的生物学效应。
采用阳离子脂质体介导的ASODN转染食管癌细胞。运用逆转录-聚合酶链反应(RT-PCR)、蛋白质免疫印迹法(Western blot)、噻唑蓝比色法(MTT法)、流式细胞术及原位凋亡细胞检测(TUNEL检测)系统研究转染细胞在体内外的生物学效应。
MTT法显示,ASODN组食管癌细胞增殖较对照组显著降低(P<0.05),Bcl-XL mRNA抑制率达57.3%,Bcl-XL蛋白显著减少,流式细胞术及TUNEL检测凋亡率分别为(31.1±5.8)%和35.0%(与对照组相比,P<0.01)。ASODN组裸鼠人食管癌生长也显著受抑(P<0.05),Bcl-XL mRNA和蛋白表达显著降低,裸鼠肿瘤细胞凋亡增加。
Bcl-XL ASODN可有效抑制食管癌细胞体外增殖及体内肿瘤生长。ASODN抑制Bcl-XL表达可为食管癌基因治疗提供治疗方法及重要理论基础。